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@#[摘 要] 目的：构建靶向CD70分子的重组免疫毒素，通过表达、纯化制备PE38与抗CD70纳米抗体重组蛋白，体外抗肿瘤实验探究重组蛋白是否对高表达CD70分子的阳性肿瘤细胞具有杀伤活性。方法：通过基因工程手段，将CD70纳米抗体Nb 2B3基因片段通过一个连接子与pET21a-PE38基因片段相连，获得重组表达载体pET21a-Nb 2B3-PE38并转入BL21（DE3）感受态细胞中进行表达、纯化与鉴定。用间接ELISA及FACS法检测Nb 2B3-PE38与CD70分子的结合活性，MTT法检测Nb 2B3-PE38对高表达CD70分子的肾透明细胞癌786-O细胞的体外杀伤活性，Annexin Ⅴ-FITC/PI双染法检测Nb 2B3-PE38对786-O细胞凋亡的影响。结果：成功构建抗CD70纳米抗体重组免疫毒素Nb 2B3-PE38，纯化获得纯度>90%的重组蛋白，SDS-PAGE及WB检测结果表明目的蛋白正确表达，分子量为56 000。纯化后的Nb 2B3-PE38能与重组CD70抗原及786-O细胞表面的CD70分子特异性结合；25 µg/mL Nb 2B3-PE38即对786-O细胞产生极显著的杀伤作用（P<0.001），并且促进786-O细胞的细胞凋亡（P<0.01），其杀伤效应强于阳性对照顺铂（P<0.01）。结论：成功制备了特异性靶向CD70分子的免疫毒素Nb 2B3-PE38，其能够有效杀伤786-O细胞并诱导细胞凋亡且效果强于顺铂。
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ABSTRACT Introduction Basketball is a sport with strong rivalry, being, therefore, a modality with great facility to cause sports injuries along its practice. Objective Investigate the status of sports injuries in college basketball for amateur students. Methods 480 college students who were not basketball experts were selected as research volunteers. The current status of sports injuries in the 12-week optional basketball course was investigated, and the main risk factors were analyzed. Results Among amateur male college students, the average playing age of third-grade male college students is higher than that of second-grade male college students, and the weekly playing frequency is lower than that of second-grade male college students. The incidence of basketball sports injuries among male college students who do not specialize in basketball from childhood to adulthood is up to 90.7%, and 85.5% during their college years. Conclusion In basketball sports, the most vulnerable parts of amateur male college students are the wrist joint, ankle joint, knee joint, leg, head and face, shoulder, waist, and back, respectively. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.
RESUMO Introdução O basquetebol é um esporte com forte rivalidade, sendo por isso uma modalidade com muita facilidade de causar lesões esportivas ao longo de sua prática. Objetivo Investigar a situação das lesões esportivas no basquetebol universitário para estudantes amadores. Métodos 480 estudantes universitários que não eram especializados em basquetebol foram selecionados como voluntários de pesquisa, e a situação atual das lesões esportivas no curso opcional de basquetebol de 12 semanas foi investigada, onde os principais fatores de risco foram analisados. Resultados Entre os estudantes universitários masculinos amadores, a idade média de jogo dos estudantes universitários masculinos da terceira série é maior do que a da segunda série, e a frequência semanal de jogo é menor do que a dos estudantes universitários masculinos da segunda série. A incidência de lesões esportivas de basquetebol entre os estudantes universitários masculinos que não são especializados em basquetebol desde a infância até a idade adulta é de até 90,7%, e 85,5% durante seus anos universitários. Conclusão No processo dos esportes de basquetebol, as partes mais vulneráveis dos estudantes universitários masculinos amadores são a articulação do pulso, articulação do tornozelo, articulação do joelho, perna, cabeça e rosto, ombro, cintura e costas, respectivamente. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.
RESUMEN Introducción El baloncesto es un deporte de fuerte rivalidad, siendo por tanto una modalidad con gran facilidad para provocar lesiones deportivas a lo largo de su práctica. Objetivo Investigar la situación de las lesiones deportivas en el baloncesto universitario para estudiantes aficionados. Métodos Se seleccionaron 480 estudiantes universitarios no especializados en baloncesto como voluntarios para la investigación, y se investigó la situación actual de las lesiones deportivas en el curso optativo de baloncesto de 12 semanas, donde se analizaron los principales factores de riesgo. Resultados Entre los estudiantes universitarios aficionados de sexo masculino, la edad media de juego de los estudiantes universitarios de tercer grado es superior a la de los estudiantes universitarios de segundo grado, y la frecuencia semanal de juego es inferior a la de los estudiantes universitarios de segundo grado. La incidencia de las lesiones deportivas de baloncesto entre los estudiantes universitarios de sexo masculino no especializados en baloncesto desde la infancia hasta la edad adulta es de hasta el 90,7%, y del 85,5% durante sus años universitarios. Conclusión En el proceso de los deportes de baloncesto, las partes más vulnerables de los estudiantes universitarios aficionados varones son la articulación de la muñeca, la articulación del tobillo, la articulación de la rodilla, la pierna, la cabeza y la cara, el hombro, la cintura y la espalda, respectivamente. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.
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@#Objective To evaluate the effect of tyrosine kinase inhibitor BGJ398 on the proliferation,apoptosis and migration of human hepatocellular cancer Huh-7 cells and explore the mechanism.Methods The effects of 10 tyrosine kinase inhibitors on the survival of Huh-7 cells were detected by MTT assay,and the sensitivity of Huh-7 cells to BGJ398 was analyzed by single-target kinetic equation and biphasic kinetic equation respectively.Huh-7 cells were added with 10,30 and 90 nmol/L BGJ398 respectively,and the control group(without drugs)was set.The effects of BGJ398 on the apoptosis and cell cycle of Huh-7 cells were detected by flow cytometry after culturing at 37℃for 24 h,the effect on the migration ability was detected by wound healing assay and the effect on the expression of multiple pathway-related proteins was detected by Western blot.Results All of 10 tyrosine kinase inhibitors inhibited the proliferation of Huh-7 cells,among which Huh-7 cells were most sensitive to BGJ398 and the IC_(50)was(0.020±0.013)μmol/L;The response of Huh-7 cells to BGJ398 was composed of two phases with F_1 accounted for 92.8%(K_(d1)was 36 nmol/L)and F_2 accounted for 7.2%(K_(d2)>1 000μmol/L).Compared with the control group,the apoptosis rate and the percentage of Huh-7 cells in G1 phase increased significantly in 30 and 90 nmol/L BGJ398 groups(t=-6.407～-4.459,each P<0.05),while the percentage of Huh-7 cells in S phase decreased significantly in 10,30 and 90 nmol/L BGJ398 groups(t=2.982,7.859 and 12.425,respectively,each P<0.05);After 24 and 48 h of scratching,the scratch area of 30 and 90 nmol/L BGJ398groups decreased significantly(t=5.376～18.197,each P<0.05);The expression levels of phosphorylated fibroblast growth factor receptor(FGFR)and phosphorylated extracellular signal-regulated kinase 1/2(Erk1/2)protein decreased significantly in 30 and 90 nmol/L BGJ398 groups(t=4.015～6.729,each P<0.01).Conclusion BGJ398 can inhibit the proliferation and migration of human hepatocellular cancer Huh-7 cells,induce apoptosis and cell cycle arrest,which might be achieved by inhibiting FGFR phosphorylation and MAPK signaling pathway.BGJ398 is expected to be a potential agent for the treatment of hepatocellular cancer.
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@#Objective To investigate the effect of a multi-target protein tyrosine kinase inhibitor,Ponatinib,on proliferation,homogeneity adhesion and migration ability of human liver cancer cell line SK-Hep-1.Methods SK-Hep-1 cells were cultured routinely and added with 24 tyrosine kinase inhibitors such as Ponatinib respectively,and the effect of Ponatinib on the survival and proliferation of SK-Hep-1 cells was detected by MTT assay.SK-Hep-1 cells were cultured routinely until the fusion degree reached 90%,then added with 0.1,0.5 and 1.0 μmol/L Ponatinib respectively,and the control group(without Ponatinib) was set up.The effect of Ponatinib on adhesion ability of SK-Hep-1 cells was detected by cell slow aggregation assay and dissociation assay,while the effect on migration ability by scratch test,and the effect on E-cadherin protein expression in SK-Hep-1 cells by Western blot.Results All 24 tyrosine kinase inhibitors inhibited SK-Hep-1 cells,among which Ponatinib showed the strongest inhibitory effect with a IC_(50) of(0.288±0.044) μmol/L.Compared with the control group,the number of cell mass(t=16.143,44.002 and 44.853 respectively,each P <0.001) and N_(TC)/N_(TE) [ratio of single cell number(N) after digestion by trypsin containing EDTA(TE) and CaCl_2(TC)](t=4.276,10.625 and 27.571 respectively,each P <0.05) decreased significantly and E-cadherin protein expression increased significantly(t=-3.757,-4.561and-6.922 respectively,each P <0.05) in 0.1,0.5 and 1.0 μmol/L Ponatinib groups;Scratch migration rate significantly decreased in 0.5 and 1.0 μmol/L Ponatinib groups(t=6.272～16.733 respectively,each P <0.01),while there was no significant difference in 0.1 μmol/L Ponatinib group(t=0.473 and 0.872 respectively,each P> 0.05) after 24 h and 48 h of scratch.Conclusion Ponatinib inhibited proliferation and migration of SK-Hep-1 cells and promoted cell adhesion.
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XueBiJing is an intravenous five-herb injection used to treat sepsis in China.The study aimed to develop a liquid chromatography-tandem mass spectrometry(LC-MS/MS)-or liquid chromatography-ultraviolet(LC-UV)-based assay for quality evaluation of XueBiJing.Assay development involved identifying marker constituents to make the assay therapeutically relevant and building a reliable one-point cali-brator for monitoring the various analytes in parallel.Nine marker constituents from the five herbs were selected based on XueBiJing's chemical composition,pharmacokinetics,and pharmacodynamics.A selectivity test(for"similarity of response")was developed to identify and minimize interference by non-target constituents.Then,an intercept test was developed to fulfill"linearity through zero"for each analyte(absolute ratio of intercept to C response,＜2％).Using the newly developed assays,we analyzed samples from 33 batches of XueBiJing,manufactured over three years,and found small batch-to-batch variability in contents of the marker constituents(4.1％-14.8％),except for senkyunolide I(26.5％).
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OBJECTIVE: The most commonly used intramedullary fixation systems for the treatment of elderly intertrochanteric fractures include the proximal femoral anti-rotation intramedullary nail (PFNA), the proximal femoral anti-rotation intramedullary nail of Asian (PFNA-II) and interlocking intramedullary nail for proximal femur (Inter-TAN), but the clinical effect of the three in the treatment of A2 and A3 type intertrochanteric fractures is still controversial. Meta-analysis was used to compare the clinical efficacy of Inter-TAN, PFNA, and PFNA-II in the treatment of unstable intertrochanteric fractures in the elderly. METHODS: A computer was used to search databases such as Cochrane, Embase, PubMed, Sinomed, CNKI, Wanfang, and VIP to collect and compare the literatures of Inter-TAN, PFNA, and PFNA-II in the treatment of elderly unstable intertrochanteric fractures. The search period was from the establishment of the database to December 2019. Two reviewers independently read the screening literature, extracted data and evaluated the quality. Meta-analysis was performed using RevMan5.3 software. RESULTS: (1) A total of eight studies were included, with 1 349 patients, including two randomized controlled studies and six cohort studies. The evidence level was ≥ III. (2) Meta-analysis showed that the operation time and intraoperative fluoroscopy time were longer, and intraoperative blood loss was more in the Inter-TAN group than those of the PFNA-II group [MD=-16.60, 95%CI (-23.22,-9.99), P 0.05). There was no significant difference in tip-apex distance, length of hospital stay, and Harris score of Inter-TAN group compared with PFNA group and PFNA-II group (P > 0.05). The incidence of postoperative screw cut-out in the Inter-TAN group was less than in the PFNA group and the PFNA-II group [OR=6.47, 95%CI(2.79, 15.00), P 0.05). CONCLUSION: For patients with osteoporosis and poor stability, Inter-TAN can provide better stability and reduce postoperative complications. For patients with poor general conditions and greater surgical risk, PFNA-II internal fixation can be selected.
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BACKGROUND: With the research and development of bone tissue engineering, it has been found that advanced glycation end products can accumulate in bone tissue and affect the structure and biomechanical properties of bone. At present, many researchers have discovered that advanced glycation end products/receptor for advanced glycation end products can induce pathological changes of osteoblasts, osteoclasts and osteocytes through special mechanisms, thereby leading to imbalance of bone reconstruction, decrease of bone strength and increase of fracture incidence. OBJECTIVE: To review the effects of advanced glycation end products on bone biomechanics and the mechanism of advanced glycation end products/receptor for advanced glycation end products on bone tissue cells. METHODS: The first author searched the relevant articles regarding the effect of advanced glycation end products/receptor for advanced glycation end products on metabolism of bone tissue cells published in PubMed, Web of Science and Medline database from January 2005 to July 2019. The results were limited to English literatures. RESULTS AND CONCLUSION: Finally, 54 representative literatures were selected for summary. The effects of advanced glycation end products on collagen cross-linking can significantly reduce bone strength. Advanced glycation end products/receptor for advanced glycation end products affects bone metabolism through pathological mechanism changes of bone tissue cells, which results in essential changes of bone tissue cells. Finally, it will lead to imbalance of bone metabolism and increase of bone fragility. The osteoporosis is directly related to the activity change of bone tissue cells, but the specific mechanism needs further study. The change of this special mechanism may provide a unique pathological mechanism, diagnosis methods, treatment and prevention strategies for osteoporosis in the future.
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BACKGROUND: The Wnt/β-catenin signaling pathway plays a key role in osteosarcoma tumorigenesis and bone remodeling, and its roles in angiogenesis and immune surveillance are the two key mechanisms of metastatic dissemination. OBJECTIVE: To review the role of Wnt/β-catenin pathway in the occurrence and development of osteosarcoma and its regulatory role in specific microenvironment. METHODS: PubMed database was searched for relevant articles on the relationship between Wnt/β-catenin pathway and bone tumors using the search terms of “Wnt/β-catenin; bone tumors.” The search time for publications was from 2000 to 2020. RESULTS AND CONCLUSION: The typical Wnt/β-catenin signaling pathway plays an important role in the growth and metastasis of osteosarcoma by directly acting on osteosarcoma cells and indirectly regulating cell activity in osteosarcoma. The Wnt/β-catenin pathway also participates in the hijacking of the bone tumor microenvironment by osteosarcoma cells, promotes the regulation of extracellular matrix, makes the tumor cells invasive, induces immune tolerance and angiogenesis in tumor cells, and eventually augments metastatic spread. In addition, the typical Wnt signaling pathway abnormally activated in osteosarcoma cells seems to be closely related to cell proliferation, induction of epithelial-mesenchymal transition-like processes, acquisition of osteosarcoma-promoting tumor cell stem cell characteristics, and metastatic spread.
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OBJECTIVE@#Abnormalities of liver-related indices are common in ICU patients, but the effects of cholestasis and hypoxic hepatitis in critically ill patients remains unclarified. The purpose of this study was to investigate the effects of cholestasis and hypoxic liver dysfunction on the prognosis of ICU patients.@*METHODS@#A retrospective study was conducted based on the data of patients admitted to the ICU for the first time between 2001 and 2011 archived in the MIMIC-Ⅲ database. The patients were divided into cholestasis, hypoxic hepatitis and control groups, and their 28-day case fatality rate as the primary outcome was compared among the groups.@*RESULTS@#A total of 5852 ICU patients were included in the analysis. The incidence of cholestasis and hypoxic liver dysfunction was 31.9% (1869/5852) and 17.9% (1046/5852), respectively. There was no significant difference in 28-day case fatality rate between cholestasis group and the control group. Compared with the control group, the patients with hypoxic hepatitis had a significantly higher 28-day case fatality rate (46% 35%, < 0.01), a higher hospital case fatality rate (40% 31%, < 0.01), and a higher ICU case fatality rate (35.7% 22.2%, < 0.01). Logistic regression analysis showed that lactic acid (LAC), aspartate transaminase (AST), and international standard ratio (INR) were independent risk factors for 28-day case fatality rate.@*CONCLUSIONS@#The incidence of cholestatic liver dysfunction is higher than that of hypoxic hepatitis, but it does not increase the 28-day case fatality rate of the ICU patients, suggesting that cholestatic liver dysfunction may be the early adaptation of the liver to critical diseases.
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Humans , Cholestasis , Hepatitis , Intensive Care Units , Prognosis , Retrospective StudiesABSTRACT
@#When the root canal is filled, the use of the adhesive resin-based sealant to form a strong bond with the dentin reduces microleakage of the apex, enhances the bending resistance of the tooth, and resists the mechanical damage of the pile preparation, thus improving the success rate of root canal treatment. However, during root canal treatment, the use of a rinsing agent may change the tissue structure of the dentin and the wettability of the dentin, thereby affecting the bonding strength between the resin sealing agent and the dentin. In this paper, the effects of a traditional root canal irrigant and several new rinsing agents on the bonding strength of resin sealant and dentin are reviewed. The results show that the traditional root canal rinsing agent NaClO is reduced as the final rinsing agent. The bonding strength of resin sealant to dentin; EDTA affects the biomechanical properties of root dentin, and the removal efficiency is low, which will affect the bond strength between the resin sealant and dentin. EDTA combined with NaClO as a final rinsing agent can improve the bonding strength. Chlorhexidine can also increase the bonding strength between the blocking agent and dentin and can be used as the final rinsing agent. The new rinsing agent MTAD has a negative impact on the resin sealing agent and dentin bond strength. QMix and maleic acid remove smear efficiency with dentin bond strengths greater than or equal to EDTA and can replace EDTA as the final rinse.
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It is increasingly evident that the microenvironment of bone can influence cancer phenotype in many ways that favor growth in bone. CD147, a transmembrane protein of the immunoglobulin (Ig) superfamily, was identified independently in different species and has many designations across different species. However, expression levels of CD147 mRNA in bone cancer have not been described. In this study, we have used real-time fluorescence quantification (RT-PCR) to demonstrate CD147 expression in malignant bone cancer and benign bone tumor tissues. The results suggested that the expression of CD147 gene was significantly up-regulated in malignant bone cancer. Moreover, we found that over-expressed RANKL progressively enhanced osteoclast formation up to 48 h, which suggested that RANKL could promote the formation of osteoclast, indicating that both CD147 and RANKL play important roles in the formation of osteoclasts. Furthermore, the expressions of four osteoclast specific expression genes, including TRACP, MMP-2, MMP-9 and c-Src, were analyzed using RT-PCR. The results indicated that four osteoclast-specific expression genes were detectable in all osteoclast with different treatments. However, the highest expression level of these four osteoclast-specific expression genes appears in the CD147+ RANKL group and the lowest expression level of these four osteoclast-specific expression genes appears with si-RANKL treatment. Characterization of the role of CD147 in the development of tumors should lead to a better understanding of the changes occurring at the molecular level during the development and progression of primary human bone cancer.
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Humans , Male , Female , Middle Aged , Aged , Osteoclasts/metabolism , Bone Neoplasms/genetics , Up-Regulation , Basigin/genetics , RANK Ligand/metabolism , Osteoblasts/cytology , Osteoblasts/metabolism , Bone Neoplasms/secondary , Bone Neoplasms/therapy , Gene Expression Regulation, Neoplastic , Blotting, Western , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Objective To observe the influence of Chinese medicine skin dialysis on traditional Chinese medicine (TCM) syndrome scores and microinflammation in patients with chronic renal failure(CRF). Methods A total of 180 patients with CRF were divided into groups A, B and C, 60 cases in each group. All of the 3 groups were given conventional treatment, and additionally, group A was given Chinese medicine skin dialysis, and group B was given pure water dialysis. Meanwhile, 60 healthy volunteers served as normal control group (group D), and had no any medication. The course of treatment lasted for one month. Before and after treatment, we examined the scores of TCM syndromes and the levels of 24-hour urinary protein, serum creatinine (SCr), blood urea nitrogen (BUN), estimated glomerular filtration rate (eGFR), albumin (ALB), immunoglobulin A(IgA), immunoglobulin G (IgG) , immunoglobulin M(IgM), interleukin 6 (IL-6) , interleukin 1(IL-1), hyper-sensitive C reactive protein (hs-CRP), and tumor necrosis factor alpha (TNF-α). Results After treatment, the scores of TCM syndromes and the levels of 24-hour urinary protein, SCr, BUN, eGFR, IL-6, IL-1, hs-CRP, and TNF-α in the 3 groups were decreased (P < 0.05 compared with those before treatment), and were markedly lower in group A than those in group B and group C (P<0.05); serum levels of IgA, IgG, IgM and ALB in the 3 groups were increased(P < 0.05 compared with those before treatment), and the levels in group A were significantly higher than those in group B and group C (P < 0.05). However, the levels of observation indexes did not arrive to the levels of group D(P<0.05). Conclusion Chinese medicine skin dialysis is effective on relieving the symptoms of CRF patients and enhancing the therapeutic effect through decreasing inflammatory mediators, inhibiting micro-inflammation and improving nutritional status.
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ABSTRACT Purpose To investigate whether intracavernosal injection of short hairpin RNA for IGFBP-3 could improve erectile function in streptozotocin-induced diabetic rats. Materials and methods After 12 weeks of IGFBP-3 short hairpin RNA injection treatment, intracavernous pressure responses to electrical stimulation of cavernous nerves were evaluated. The expression of IGFBP-3 and IGF-1 at mRNA and protein levels were detected by quantitative real-time PCR analysis and Western blot, respectively. The concentration of cavernous cyclic guanosine monophosphate was detected by enzyme-linked immunosorbent assay. Results At 12 weeks after intracavernous administration of IGFBP-3 shRNA, the cavernosal pressure was significantly increased in response to the cavernous nerves stimulation compared to the diabetic group (P<0.05). Cavernous IGFBP-3 expression at both mRNA and protein levels was significantly inhibited. At the same time, cavernous IGF-1 expression was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Cavernous cyclic guanosine monophosphate concentration was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Conclusions Gene transfer of IGFBP-3 shRNA could improve erectile function via the restoration of cavernous IGF-1 bioavailability and an increase of cavernous cGMP concentration in the pathogenesis of erectile dysfunction in streptozotocin-induced diabetic rats.
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Animals , Male , Penis/drug effects , Insulin-Like Growth Factor Binding Protein 3/pharmacokinetics , RNA, Small Interfering/pharmacokinetics , Diabetes Mellitus, Experimental/physiopathology , Erectile Dysfunction/physiopathology , Erectile Dysfunction/drug therapy , Insulin-Like Growth Factor I/analysis , Insulin-Like Growth Factor I/drug effects , Enzyme-Linked Immunosorbent Assay , Biological Availability , Random Allocation , Blotting, Western , Reproducibility of Results , Rats, Wistar , Streptozocin , Diabetes Mellitus, Experimental/complications , Real-Time Polymerase Chain Reaction , Erectile Dysfunction/etiology , InjectionsABSTRACT
The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg(-1)) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg(-1), i.v.) or different dosage (18.9, 37.8 and 75.6 g·kg(-1), i.g.) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg(-1), i.g.) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg(-1)), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg(-1)) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg(-1) of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g·kg(-1)) could result in liver injury in the LPS-based idiosyncratic hepatotoxicity model, which could be used to evaluate the idiosyncratic hepatotoxicity of PM.
Subject(s)
Animals , Rats , Alanine Transaminase , Blood , Aspartate Aminotransferases , Blood , Chemical and Drug Induced Liver Injury , Pathology , Hepatocytes , Pathology , Lipopolysaccharides , Polygonum , Toxicity , Rats, Sprague-DawleyABSTRACT
To investigate the difference of liver injury in rats gavaged with crude and processed Polygoni Multiflori Radix. The 75% ethanol extract of crude and processed Polygoni Multiflori Radix (50 g · kg(-1) crude medicine weight/body weight) were continuous oral administered to rats for 6 weeks. Serum biochemical indicators were dynamically detected, the change of liver histopathology was assessed 6 weeks later. Principal component analysis (PCA) was adopted to screen sensitive indicator of the liver damage induced by polygoni multiflori radix. Biochemical tests showed that the crude Polygoni Multiflori Radix group had significant increase of serum ALT, AST, ALP, DBIL and TBIL (P < 0.01 or P < 0.05) and significant decreases of serum IBIL and TBA (P < 0.01 or P < 0.05), while the processed Polygoni Multiflori Radix group showed no obvious changes, compared to the untreated normal group. Histopathologic analysis revealed that crude Polygoni Multiflori Radix group exhibited significant inflammatory cells infiltration in portal area around the blood vessels, tissue destruction and local necrosis of liver cells. There were not obvious pathological changes in processed Polygoni Multiflori Radix group. The results demonstrated that the injury effect of processed Polygoni Multiflori Radix on liver injury of rats was significantly lower than that of unprocessed, and that processing can effectively reduce the hepatotoxicity of Polygoni Multiflori Radix. Traditional transaminase liver function indicators were not sensitive for crude Polygoni Multiflori Radix induced liver damage. The serum content of DBIL and TBIL can reflect the liver damage induced by crude Polygoni Multiflori Radix early and can be sensitive indicators for clinical monitoring the usage of it.
Subject(s)
Animals , Female , Male , Rats , Chemical and Drug Induced Liver Injury , Chemistry, Pharmaceutical , Methods , Drugs, Chinese Herbal , Chemistry , Toxicity , Liver , Wounds and Injuries , Plant Roots , Chemistry , Toxicity , Polygonum , Chemistry , ToxicityABSTRACT
Background A simple, rapid, low-cost and environmentally friendly method was developed to determine dopamine (DA) in the presence of ascorbic (AA) and uric acid (UA) based on a novel technique to prepare a graphene-chitosan (GR-CS) nanocomposite and modify it on the surface of carbon paste electrode (CPE). For our design, CS acts as a media to disperse and stabilize GR, and then GR plays a key role to selective and sensitive determination of DA. Results Under physiological conditions, the linear range for dopamine was determined from 1 × 10- 4 to 2 × 10- 7 mol/L with a good correlation coefficient of 0.9961 in the presence of 1000-fold interference of AA and UA. The detection limit was estimated to be 9.82 × 10- 8 mol/L (S/N = 3). In order to study the stability and reproducibility, GR/CS/CPE underwent successive measurements in 10 times and then tested once a d for 30 d. The result exhibited 98.25% and 91.62% activities compared with the original peak current after 10-time measurements and 30-d storage. Conclusion The GR/CS/CPE has wide linear concentration range, low detection limit, and good reproducibility and stability, which suggests that our investigations provide a promising alternative for clinic DA determination.
Subject(s)
Carbon/chemistry , Dopamine/analysis , Chitosan/chemistry , Graphite/chemistry , Electrodes , Nanocomposites/chemistry , Electrochemical Techniques , Limit of Detection , Chemistry Techniques, Synthetic , Hydrogen-Ion ConcentrationABSTRACT
Background: Light can be absorbed by bacterial pigment and affects its growth. Prodigiosin is a red pigment found in various bacterial species. The purpose of this study was to investigate the impacts of light on prodigiosin production, biomass formation, and membrane integrity of Serratia marcescens y2. Results: S. marcescens y2 grew better and produced more intracellular prodigiosin in darkness than in illumination. The pigment leakage ratio from cells was detected more in light than in darkness conditions. Ethidium bromide uptake assay could visually prove the prodigiosin-related loss of membrane integrity under illumination. A higher concentration of malondialdehyde (MDA) was detected in light-treated culture than in darkness. Tests of different light treatments (red, yellow, blue and green) showed that the maximum extracellular pigment and the minimum biomass formation and intracellular pigment were obtained in green light. Conclusions: Prodigiosin could absorb light, and then initiate phototoxicity damage of the cytomembrane.
Subject(s)
Prodigiosin , Serratia marcescens/metabolism , Malondialdehyde/analysis , Lighting , Chromatography, High Pressure Liquid , Biomass , EthidiumABSTRACT
<p><b>OBJECTIVE</b>To construct the zinc finger protein-activating transcription factor (ZFP-ATF) plasmid and evaluate its efficacy in inducing vascular endothelial growth factor (VEGF) expression in EY.HY926 endothelial cells.</p><p><b>METHODS</b>Firstly, we constructed the ZFP-ATF plasmid, then testified the quantity of VEGF protein in EY.HY926 endothelial cells after transfected with ZFP-ATP plasmid by Western blot, finally, we used the RT-PCR to testify whether the ZFP-ATF can stimulate expression of VEGF splice variants.</p><p><b>RESULTS</b>The ZFP-ATF DNA sequences were located the multiclone sites of PVAX1 vector between the site of BamH1and Xhol.Western blot result showed VEGF expression in EY.HY926 endothelial cells transfected with ZFP-ATF plasmid was significantly higher than that in cells transfected with VEGF165 (19.95±3.95 vs.12.15±1.55 μg÷μL, P<0.01).RT-PCR result showed VEGF-A mRNA expression level induced by ZFP-ATF was high than that induced by VEGF165.</p><p><b>CONCLUSION</b>ZFP-ATF can up-regulate the VEGF-A expression in comparison with VEGF165, which might have beneficial effects in angiogenesis process.</p>
Subject(s)
Humans , Activating Transcription Factors , Physiology , Amino Acid Sequence , Base Sequence , Cells, Cultured , Endothelial Cells , Metabolism , Molecular Sequence Data , Neovascularization, Physiologic , Plasmids , Up-Regulation , Vascular Endothelial Growth Factor A , Genetics , Zinc Fingers , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the role of multidetector row spiral computed tomography in the diagnosis of popliteal artery entrapment syndrome (PAES).</p><p><b>METHOD</b>Ten patients (13 limbs) were diagnosed as PAES through open surgery in Peking Union Medical College Hospital from January 2006 to December 2009, among whom three (30.0%) had both limbs involved.</p><p><b>RESULTS</b>Eleven limbs (84.6%) had occlusions in popliteal artery at admission. While only 15.4% of the patients were diagnosed as PAES by positional stress test, Doppler ultrasound, and arterial angiography, the disease was confirmed in all 10 patients by spiral computed tomography. As shown by inter-operative exploration, the PAES types included type 1 (n=2, 15.4%), 2(n=5, 38.5%),3 (n=4, 30.8%), and 5 (n=2, 15.4%). Two limbs underwent popliteal artery release only, 8 limbs underwent politeal artery release, thromboendarterectomy, and angioplasty with a venous or prosthetic patch, and 3 patients underwent catheter-based thrombolysis and popliteal artery release subsequently. During the follow-up (mean: 22 months; range: 4-33 months), the primary patency rate was 92.3% (12/13), the total patency rate was 100% (13/13), and the limb salvage rate was 100%.</p><p><b>CONCLUSION</b>Positional stress test, Doppler ultrasound, and arterial angiography play limited roles in the diagnosis of PAES with popliteal artery occlusion, while multidetector row spiral computed tomography is much helpful in diagnosing and typing PAES.</p>